We are IntechOpen,

the world’s leading publisher of

Open Access books
Built by scientists, for scientists

4,200
Open access books available
116,000
International authors and editors
125M
Downloads

Our authors are among the

154
Countries delivered to
TOP 1%
most cited scientists
12.2%
Contributors from top 500 universities

Selection of our books indexed in the Book Citation Index

in Web of Science™ Core Collection (BKCI)

Interested in publishing with us?

Contact book.department@intechopen.com
Numbers displayed above are based on latest data collected.
For more information visit www.intechopen.com
 5

Establishment of Functional Biotechnology

Laboratories in Developing Countries
Marian D. Quain, James Y. Asibuo,
Ruth N. Prempeh and Elizabeth Y. Parkes
Council for Scientific and Industrial Research, Crops Research Institute, Kumasi,
Ghana

1. Introduction
Traditionally, biotechnology is defined as making use of living organisms or genetic
material from living organisms to provide new products for agricultural, industrial, and
medical uses. This definition includes the use of fermentation in the leavening in the 10000
BC. This technology over the years has advanced into Modern Biotechnology. According to
the Cartagena protocol (Secretariat of the Convention on Biological Diversity, 2000),
Biotechnology is defined as any technological application that uses biological systems, living
organisms, or derivatives thereof, to make or modify products or process for a specific use.
According to the Convention on Biological Diversity, Art.3 (i), “Modern biotechnology”
means the application of:
a. In vitro nucleic acid techniques, including recombinant deoxyribonucleic acid (DNA)
and direct injection of nucleic acid into cells or organelles, or
b. Fusion of cells beyond the taxonomic family, that overcome natural physiological
reproductive or recombination barriers and that are not techniques used in traditional
breeding and selection.
Plant Biotechnology encompasses tools such as tissue culture and molecular biology which
are used in crop improvement. Although these technological tools are applied in advanced
countries, their use in agricultural research and development in developing countries is
limited. However, these countries need to enhance the utilization of tissue culture and
molecular biology to increase agriculture productivity. The prospects of biotechnology as a
modern tool for addressing various productivity problems and challenges in agriculture in
the face of present day changing climatic conditions and starvation are now well known.
Agriculture accounts for about 40% of Ghana’s GDP, contributes 35% of foreign exchange
earnings, and provides employment for over 60% of the population. More than 80% of the
rural populations depend on it for their livelihood.
In recognition of the need to use biotechnology tools in agriculture in sub-Sahara Africa, in
June 2003 at a Worldwide Ministerial Conference, in Sacramento, USA 112 Ministers of
Science and Technology from 117 countries recommended the facilitation of access of
Developing countries to Science and Technology innovations as means to reach Millennium

www.intechopen.com
66 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

development goals (MDGs). As a follow-up to that, in June 2004, a West African Ministerial
Conference on “the use of Science and Technology to improve agricultural productivity in
Africa” was held in Ouagadougou (Burkina Faso). At this meeting, participants recognised
the need to:

• Develop an information strategy on new technologies and especially on biotechnology

• Establish a strong scientific partnership between research institutions of Africa and
developed countries
• Put in place a Regional Biotechnology Centre
Subsequently, in November 2004, there was the Economic Community of West African
States (ECOWAS) Ministerial Conference on “Agriculture and Biotechnology” in Abuja
(Nigeria) the meeting recommended the following:
• The need to establish Regional Biotechnology Centres of Excellence in countries having
comparative advantages
• The promotion of in situ Research and Development activities on priority areas to
support the emergence and growth of the biotechnology industry in West Africa
• The transfer of biotechnology product packages and their commercialisation in relevant
areas
• The reinforcement of private-public sectors collaboration in order to boost the local
Biotechnology industry
• The reinforcement of regional and national capacities in Biosafety
There was thus adoption of the Biotechnology and Biosafety Programme (BBP) Action Plan
by the ECOWAS Ministerial Conference in Accra (Ghana), in May 2007. The BBP general
objective is to use the development and exploitation of biotechnology products as means to
increase agricultural productivity and competitiveness in West Africa
The Specific Objectives are to:
1. Promote the use of Biotechnology in agriculture
2. Develop a regional approach for Biosafety
3. Establish and make effective at the regional level, a mechanism for coordination of
initiatives, fund raising and communication in the field of Biotechnology and Biosafety.
Thematic areas addressed in priority being:
• The application of Molecular Markers
• The application of Genetic Engineering
• The application of Molecular Diagnostics for animal and plant diseases
• Plant tissue/cell culture and micro-propagation techniques
• Vaccines for livestock production
• Animal Reproduction Technologies
This meeting was a significant landmark in the application of Science and Technology for
improving performance in the agribusiness sector in the West African sub-region. This is
because the meeting brought together biotechnology and biosafety experts, as well as the
ECOWAS Ministers of Environment, Science and Technology, and Agriculture. At the end
of the meeting, the communiqué issued indicated that the Ministers fully support the

www.intechopen.com
Establishment of Functional Biotechnology Laboratories in Developing Countries 67

application of biotechnology in addressing some of the numerous problems facing

agriculture in Africa, particularly towards improvement in production, competitiveness and
sustainable management of natural resources. Although the Ministers pledged their
support, they stressed the need to have in place safety measures to ensure effective and
sustainable application of the technologies.
All these initiatives have harnessed the application of modern techniques, however, in
Ghana, utilization of biotechnology tools in agricultural research and development is
associated with several setbacks: which may lead to nonfunctional and unsustainable
laboratories. This paper thus focuses on how functional laboratories have been established
in Ghana, with specific reference to some research output by the Biotechnology Unit of the
Council for Scientific and Industrial Research (CSIR) – Crops Research Institute (CRI).

2. Establishment of biotechnology laboratories and some research output

2.1 Tissue culture
Tissue culture is the most applied biotechnology tool and its establishment is vital for the
application of various techniques. By means of definition, plant tissue culture is the growth
or maintenance of plant cells, tissues or organs or whole plant on a nutrient culture medium
under aseptic conditins in vitro. Tissue culture employs the principle of "Totipotency”
which is the cell characterestics in which the potential for forming all the cell types in the
adult organsim is retained, or the capacity of differentiated cells to retain their full genetic
potentialities and express them under appropriate conditions, or potential of cells or tissues
to form all cell types and/ or to regenerate plants (Murch & Saxena 2005).
Listed below are applications and advantages of plant tissue culture
• Production of clones
• Large-scale plant multiplication
• Mutation-assisted breeding
• Induction of genetic variability- somaclonal variation
• In vitro selection
• International germplasm exchange
• All year round availability of tissue culture derived plants
• High commercial prospects – floriculture and vegetative crops
• Plants as a bioreactor for producing vaccines, and chemicals
• Saves time and space
• Long-term storage of elite genetic material
• Establishment of germplasm bank
• Labour oriented- employment generation, socio-economic impact
• Secondary metabolite production- medicine
Plant tissue culture techniques include:
• Anther/pollen/microspore/ovary culture
• Protoplasts
• Embryo rescue
• In vitro fertilization

www.intechopen.com
68 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

• In vitro micro-grafting
• Micropropagation
• Somatic embryogenesis
• Callus and cell suspension
• Cryopreservation
• Cold storage
• Encapsulation
• Bioreactor
• Gene transfer
In Ghana, one of the first agricultural based research organizations to set up a tissue culture
laboratory was the Ghana Atomic Energy Commission (GAEC) in the mid 1980s. This was
followed by the Department of Botany of the University of Ghana, Legon in 1988, to train
students. Subsequently, the Council for Scientific and Industrial Research (CSIR) Crops
Research Institute (CRI) also established a tissue culture laboratory in 1996. All these set ups
had to cope with interrupted supply of water and electricity, however, putting in place
efficient water storage systems as well as bore hole and also standby generator for electricity
helped solve these problems. Other setbacks included lack of regular and reliable source of
consumables, glassware, equipment, equipment maintenance, not to mention source of funds,
since limited funds were received from central government. However, it is worth mentioning
that the CGIAR centers have been instrumental in training human resources and assisting with
supplies through projects. The Consultative Group on International Agricultural Research
(CGIAR) centers include the International Institute for Tropical Agriculture (IITA),
International Center for Tropical Agriculture (CIAT), International Crops Research Institute
for the Semi-Arid-Tropics (ICRISAT), just to mention a few.
When setting up the tissue culture facility of the CSIR-CRI, the laboratory initially used to
share laminar flow cabinet with the microbiology research group. This was very frustrating
since it took us a while to establish clean cultures. Once we established our ability to
produce results in tissue culture, the laboratory in 1998 collaborated in research activities
sponsored by German Technical Cooperation (GTZ), West Africa seed Development Unit
(WASDU) for the production of clean plating materials of yam and cassava in West Africa
(Quain, 2001). Another project with IITA with funding from GATSBY UK, produced clean
Musa planting material for field evaluation and selection of hybrids with tolerance to the
Black Sigatoka Disease in Ghana which started in 1998 this project resulted in the selection
an release of two hybrid Musa species for release and utilization in Ghana. Also, in 1999, as
the institute’s sweetpotato breeding group worked towards the selection of varieties to be
released to farmers under the Root and Tuber Improvement Project (RTIP), the laboratory
with assistance from IITA, produced clean planting materials for multilocational trial. The
sweetpotato varieties cleaned through tissue culture techniques in the laboratory, when
established in the field was highly accepted by Agriculture extension officers and farmers.
The tuber yield resulted in a 30% increase when compared with the conventional planting
material (Otoo & Quain 2001).
Subsequently, the CSIR-CRI tissue culture laboratory has optimized existing protocols for
local crop varieties, some of these recent research outputs include publications on the
following: Multiple Shoot Generation Media for Musa sapientum L. (False Horn,
Intermediate French Plantain and Hybrid Tetraploid French Plantain) Cultivars in Ghana

www.intechopen.com
Establishment of Functional Biotechnology Laboratories in Developing Countries 69

(Quain et al., 2010a). This paper considered plantains (Musa sapientum), a major staple in
Ghana, which encounter several production constraints including availability of adequate
healthy planting materials at the time the crop needs to be planted. In attempts to improve
production, tissue culture methods were employed, using one medium. It was however
realized that optimization of invitro rapid propagation protocol for mass production of
different accessions of Musa was paramount. Excised buds from cultures with proliferating
buds were used as explants in this experiment. The cultures of proliferating buds had been
generated from excised apical meristem of four Musa varieties (False Horn; local names –
Osoboaso and Apantu, intermediate French plantain; local name – Oniaba and FHIA 21,
which is a hybrid tetraploid French plantain) which were cultured on Murashige and Skoog
(MS) medium (Murashige and Skoog, 1962) containing indole-3-acetic acid (IAA), citric acid,
and 0-20 μM benzyl amino purine (BAP). The most popular local plantain variety, Apantu,
only produced proliferating buds profusely when placed on routine medium (MS medium
containing IAA, citric acid and 20 μM BAP). Reducing the concentration of BAP generated
an average of more than 4 shoots/culture in 8 weeks. Medium not supplemented with any
plant growth regulators also generated an average of less than 2 shoots/culture in 8 weeks.
The other three Musa varieties generated 4-8 shoots/culture from proliferating buds,
indicating that each cultivar has optimum concentrations at which rapid plantlet formation
can be optimized to meet growing demands for planting material. This protocol has
presently been adapted by the laboratory which produces about 3000 musa plants yearly
through tissue culture for farmer, NGOs and interested organizations.
Other research activities have also established in vitro manipulation protocols for Dioscorea
species (yam), which is a vital staple. In the yam tissue culture research, effect of various
hormonal (growth regulators) combinations on in vitro sprouting of various species of
Dioscorea spp under light and dark conditions (Ashun, 1991). In vitro studies on
micropropagation of various yam species (Dioscorea species) (Ashun, 1996), indicated that
where various concentrations of phytohormone Naphthalene Acetic Acid (NAA), 2,4-
dichlorophenoxycetic acid (2,4-D), and BAP are used to culture Dioscorea spp in vitro using
complete Murashige and Skoogs medium, the concentrations of 0.5µM and 5µM NAA,
enhanced plantlet development. BAP concentrations of 5µM and above were lethal to explant
development whereas 5µM and above NAA enhanced callus development in D. alata cv. 145
used. These studies also established that during yam explants initiation in culture with
explants derived from vine, the age of the explants is critical. Explants aged two to 20 weeks
were used in this study and for the different Dioscorea species used, the highest growth for D.
bulbifera was in 2 week old explants, D. dumentorum were six weeks and four week old
explants for D. alata (Quain & Achempong, 2001). Dioscorea species produce tubers and it is an
important staple in Ghana and sub-saharan African countries. The above mentioned tissue
culture studies therefore provide the basic tissue culture tools applicable in modern
biotechnology, that can be used in the improvement of the crop in the sub-region.
Current tissue culture research activities are aiming at producing protocol for the in vitro
manipulation of local bananas, plantains as well as various local root and tuber crops. The
aim of these is to establish schematic mass production systems to benefit the commercial
farmer. Protocols for long-term in vitro conservation of germplasm are also being optimized.
The development of all these protocols will facilitate the adaptation of other modern
biotechnology tools for the maintenance and improvement of local crop varieties to meet
agricultural production constraints.

www.intechopen.com
70 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

2.2 Molecular biology

Molecular biology is the aspect of biology that deals with the molecular basis of biological
activity. This aspect of science is related with other areas of biology, chemistry, genetics and
biochemistry. Mostly, molecular biology chiefly covers interactions between the various
systems of a cell, namely between the different types of DNA, RNA and protein
biosynthesis, and how these interactions are regulated. In 1961, William T. Astbury, made a
statement that “molecular biology implies not so much a technique as an approach, an
approach from the viewpoint of the so-called basic sciences with the leading idea of
searching below the large-scale manifestations of classical biology for the corresponding
molecular plan. It is concerned particularly with the forms of biological molecules and with
evolution, exploitation, and ramification of those forms of the ascent to higher and higher
levels of organisation. Molecular biology is predominantly three-dimensional and
structural—which does not mean, however, that it is merely a refinement of morphology. It
must at the same time inquire into genesis and function” (Astbury, 1961).
Molecular biologists have since the late 1950s and early 1960s learned to characterize,
isolate, and manipulate the molecular components of cells and organisms. These
components include firstly, DNA, which is the storehouse of genetic information. Secondly
is RNA, which is a close relative of DNA with functions ranging from serving as a
temporary working copy of DNA to actual structural and enzymatic functions, as well as a
functional and structural part of the translational apparatus. Thirdly, are proteins; the major
structural and enzymatic type of molecule in cells (Molecular Biology Source:
http://en.wikipedia.org/w/index.php?oldid=417169395).
The aspects of biology listed below can all be found under Molecular Biology:
• Genomics
• Proteomics
• Molecular Microbiology
• Genetic transformation
• Molecular modelling
• Molecular breeding
• Molecular marker selection
• Mutation
• Bioinformatics
• Genetic fingerprinting
Following the successful establishment of tissue culture facility at the CSIR – CRI, it became
apparent that a molecular biology laboratory be establish to complement the biotechnology
activities. Project proposals developed under the then Agricultural Services Sub-Sector
Investment Programme (AGSSIP) project secured the necessary basic equipment for
molecular biology research. The project also contributed toward the training of two
researchers; one in marker assisted breeding and another in genetic transformation in
advanced laboratories. These scientists returned to Ghana, with the basic laboratory
consumables to initiate the molecular biology laboratory. With assistance from a research
assistant who had just completed using molecular techniques in disease diagnosis in a
Ghanaian university, the molecular biology laboratory started operation in 2006 supported
with inputs from the Generation Challenge Programme (GCP), CIAT and International
Atomic Energy Agency (IAEA). In that same year, funds were also secured from the
Government of Ghana to provide more equipment and consumables for the laboratory.

www.intechopen.com
Establishment of Functional Biotechnology Laboratories in Developing Countries 71

These gave the laboratory a sound basis to be really established. Since then the CSIR-CRI
molecular biology laboratory has been locally adjudged the best biotechnology laboratory in
Ghana and is yearly conducting training courses for researchers and students both locally
and within the West African sub-region. Several research publications have been released
and these include the following:
Assessing transferability of Sweetpotato EST SSR primers to cocoyam and micropropagation
of nine elite cocoyam varieties in Ghana; Cocoyam, an important staple crop in Ghana,
provides edible leafy vegetable and starchy cormels. Due to difficulty in getting primers for
cocoyam, sweetpotato EST SSR primers were used to amplify genomic DNA of elite
cocoyam lines. Genomic DNA was isolated from 10 sweetpotato and nine cocoyam
cultivars. Ten sweetpotato accessions were screened alongside three cocoyam cultivars,
using 22 EST Sweetpotato SSR primers, 13 of which could amplify cocoyam sequences and
were subsequently used to screen nine cocoyam cultivars. Thirteen random primers were
also used for diversity study. Cocoyam cultivars were established in vitro. Dendogram
generated after screening cocoyams alongside sweetpotato, grouped sweetpotato varieties
in two main clusters and cocoyam in one cluster. The random primers and the SSRs grouped
the cocoyam into two clusters which corresponds with known morphological classification.
The method would be used to screen large cocoyam germplasm (Quain et al., 2010b).
Through this study genetic fingerprint of eight elite and one local check cocoyams has been
documented. These fingerprinted cocoyam accessions are presently being evaluated on the
field to establish their agronomic attributes and select some lines for release to farmers and
the Ghanaian public for utilization. This will be the first time ever in Ghana that research
output is releasing cocoyam varieties.
Genetic diversity of elite Musa cultivars and introduced hybrids in Ghana using SSR markers;
in this study, molecular diversity was carried out on 10 Musa cultivars using SSR. Musa SSRs
(49) marker was used for the diversity and NTSYS Data analysis used to establish conclusions
on studies. Dendogram and similarity matrix generated, indicated that local false horn and
intermediate French plantain are distantly related (16.78%). The closest related cultivars are
two false horn (Apantu-Dichotomy and Osoboaso) at 70.32%. Similarity between introduced
hybrids and local false horn plantains and local intermediate French plantains was in the range
of 20.81–49.67% and 18.85–42.27% respectively. Apem (local intermediate French plantain) was
distantly related to all the cultivars screened (16.78–36.84%). The information generated has
documented diversity between the introduced hybrids and elite local cultivars and this will
aid breeders mine for genes in the local cultivars that are responsible for earliness, peculiar
taste and preferred cooking qualities (Quain et al., 2010c).
Genetic relationships between some released and elite Ghanaian cassava cultivars based
on distance matrices has also been carried out (Acquah et al., 2010); Eleven (11) released
and two local Ghanaian cassava cultivars were fingerprinted to estimate the genetic
diversity among them using 35 SSR markers. Genomic DNA of thirteen cassava cultivars
(UCC,IFAD, Agelifiaa, Nyerikobga, Nkabom, Essam Bankye, Akosua Tumtum, Debor,
Filindiakong, Afisiafi,Doku Duade, Bankye Hemaa and Bankye Botan) were isolated and used
as template for PCR amplification involving 35 SSR markers. The recorded gel bands (163
polymorphic bands) were subjected to NTSYS Version 2.1 software for cluster analysis
and development of dendrogram to show the corresponding similarity coefficients.
Genetic relationships between Bankye Hemaa and Filindiakoh and that between Bankye
Hemaa and Afisiafi had similarity coefficients of 1.2%. The local cultivars, Debor and Akosua
Tumtum were related at 52.31% similarity. Filindiakoh was found to be the relative to

www.intechopen.com
72 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

Akosua Tumtum and Debor at 17.9 and 29.1% similarity, respectively. Bankye Botan and
Bankye Hemaa, however, were distantly related to most of the cultivars, including the local
varieties. Bankye Botan and Bankye Hemaa are distant relatives to most of the cultivars,
including the local varieties which could however make these cultivars also very useful in
breeding. This research work documented molecular information on released cassava
varieties for the first time in Ghana. This information will contribute towards variety
identification as they are released for utilization by farmers. The various research groups
that have released cassava varieties over the years can also track the performance of their
lines within the country and the subregion.
Groundnut is a member of the genus Arachis and the crop is divided into two subspecies
and six botanical varieties based on morphological characteristics. A groundnut core
collection of 831 accessions was developed from a total of 7432 US groundnut accessions
based on morphological characteristics. Identification of DNA markers associated with the
botanical varieties of groundnut would be useful in genotyping, germplasm management
and evolutionary studies. A study was initiated to evaluate 22 groundnut genotypes
representing six botanical varieties from a US groundnut core collection to determine their
diversity using DNA microsatellites. Cluster analysis located the lines in their assigned
specific botanical groups in agreement with available morphological classification for
groundnut (Asibuo et al., 2010). Groundnut production and utilization in Ghana is presently
very promising and selected groundnut varieties are produced for the confectionary
industry. The output of this study will thus enable organizations that produce seed for
farmers to cultivate scrutinize the stability and identity of their seeds.

2.3 Cryopreservation
Cryopreservation is a process of cooling cells, tissues or organs at ultralow temperature to
preserve them indefinitely. The applied temperature is typically at −196 °C which is the
boiling point of liquid nitrogen. Other temperatures applied in cryopreservation include -
40°C, -70°C, -80°C, in programmable or ultra-low freezers, or in the vapor phase of liquid
nitrogen at -150°C, or at -210°C in nitrogen slash. Technically, at these low temperatures,
any biological, metabolic activity as well as biochemical reactions that would lead to the cell
losing viability should cease and the preserved cell tissue or organ should be viable when
retrieved from cooling. Due to the complex nature of cells, organs and tissues subjected to
cryopreservation, other additives including cryoprotectants are used to prevent damage
otherwise caused by cooling.
As the biotechnology research advanced, conservation of clonally propagated crops was
identified as an important aspect. To facilitate the development of this technological tool,
a PhD student worked on Complementary Conservation of Root and Tuber Genetic
Resources - Dioscorea species and Solenostemon rotundifolius. The major focus of this study
was the application of cryopreservation techniques to complement in vitro slow growth
methods, since in vitro conservation under slow growth has been used for the
conservation of clonally propagated crops. However, it demands periodic subculturing
and regular attention and, with interruptions in electric power supply, conserved cultures
are in danger of being lost. Presently the existing root and tuber germplasm conservation
techniques serve a short to medium-term purpose only. The ultimate means of long-term
conservation, which will complement all the existing modes being used and serve the
purpose of base collections, is conservation in liquid nitrogen at –196oC (Engelmann,
2000). Storage of biological material at ultra-low temperatures, preferably that of liquid

www.intechopen.com
Establishment of Functional Biotechnology Laboratories in Developing Countries 73

nitrogen, arrests all metabolic activities: consequently, no genetic changes occur,

theoretically, permitting indefinite germplasm storage periods (Panis & Lambardi, 2005).
However, in practice, although very long, indefinite (seed) storage may not be attainable
(Walters et al., 2005) and this probably applies to other forms of germplasm as well
(Benson & Bremner, 2004). The development of protocols tested three cryomodels: being
vitrification–based, silica gel dehydration, encapsulation vitrification, as well as
encapsulation desiccation. The study revealed that, the successful cryopreservation of
Dioscorea rotundata which is an important crop producing mealy tubers is possible using a
simple vitrification protocol. The procedure incorporates:
- pregrowth of the donor plant on 0.09 M sucrose-supplemented medium for five weeks
- preculture on 0.3 M sucrose supplemented medium for 5 d
- PVS2 solution for 40 min
- Rapid cooling in liquid nitrogen or slow cooling to -80ºC
Through this study, Dioscorea rotundata accession “Pona” which is an elite variety in
Ghana was successfully cryopreserved for the first time. The technique developed was
simple, cost-effective and potentially reliable methodology that does not require
sophisticated equipment. The procedures can be adapted for germplasm conservation of
other species, using limited resources in laboratories in sub-Saharan Africa. It was also
brought to the fore that to achieve an optimal recovery of cryopreserved explants, the
donor plants should be adequately conditioned. S. rotundifolius (Frafra potato) was
extremely sensitive to the vitrification based protocol. The nodal explants used in the
experiments easily becomes hyperhydric, and are impossible to dehydrate sufficiently for
cryopreservation, this provide a sound basis for further attempts to cryopreserve Frafra
potato genetic resources. Presently, cryopreservation tools are being developed further for
other vegetatively propagated crops in Ghana.

2.4 Marker assisted breeding

Marker assisted selection (MAS) is indirect selection process where a trait of interest is
selected, not based on the trait itself, but on a marker linked to it (Ribaut and Hosington,
1998). During selection for tolerance to an abiotic or biotic stress by means of MAS, the
plants not on basis of quantified but rather a marker allelomorph (allele) which is linked
with the particular trait (disease) is used to determine the presence of the trait (disease). The
assumption being that linked allele associates with the gene and/or quantitative trait locus
(QTL) of interest. MAS can be useful for traits that are difficult to measure, exhibit low
heritability, as well as those that are expressed late in development.
Over the years most breeding activities in the CSIR-CRI have been mainly through selection,
based on agronomic traits. However, since 2005, due to training obtained from The
International Centre for Tropical Agriculture (CIAT), Cali Columbia, the cassava breeding
group has initiated development of crosses to select varieties. Some of the breeding efforts
are towards pyramiding genes responsible for tolerance/resistance to cassava mosaic
disease using wild relatives from the centre of origin in South America into local Ghanaian
accessions through mechanical hybridization. Other similar crop development to select
varieties that can withstand biotic and abiotic stresses include drought tolerance in maize,
pod shattering in soybean and disease resistance in peanuts. All these processes are being
hastened by the utilization of molecular markers to shorten the number of years used in
breeding from 10 – 15 years to about four to six years.

www.intechopen.com
74 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

2.5 Plant disease diagnostics

Effective crop management involves the early and accurate diagnosis of plant disease. Early
introduction of effective control measures in plant development can facilitate plant diseases
management. Reliance on symptoms is usually not satisfactory in this regard. This is
because the disease may be well underway when symptoms first appear, and symptom
expression can be highly variable. Biological techniques for disease diagnosis and pathogen
detection are usually highly accurate but too slow and not amenable to large-scale
application. Recent advances in molecular biology and biotechnology are being applied to
the development of rapid, specific, and sensitive tools for the detection of plant pathogens.
(Miller & Martin, 1988). Some Immunological and nucleic-acid hybridization-based methods
available for pathogen detection in crop systems are listed below:
• Immunoassay Technology
• Enzyme Linked Immunoabsorbent Assay (ELISA)
• Colloidal Gold
• Immunofluorescence Assay (IFA)
• Radio Immunoassy (RIA)
• Nucleic – Acid hybridization – Based Pathogen detection
• Nucleic Acid-Based Detection Technologies
• Dot – Blot Assay
• Nonradioactive Labels
• Restriction Fragment Length Polymorphisms (RFLPS)
• Nucleic Acid Probes
• Uncloned Probes
• Synthetic Probes
• Cloned Probes and RFLPS
• Viruses and Viroids
• Mycoplasma – like organisms and bacteria
• Fungi
• Nematode
Molecular biology tools that have been explored so far in our Laboratory include the
application of molecular markers to screen for occurrence of disease in crops. This is being
applied for African Cassava Mosaic Virus (ACMV) in cassava, Tomato Yellow Leaf Curl Virus
(TYLCV) and Root Knot diseases in tomatoes, yellow mottle virus in rice and Sweet Potato
Virus Disease (SPVD) in sweetpotato is still at the developmental stages. In a recent study on
cassava, disease observations in the field were confirmed with laboratory diagnostics using the
polymerase chain reaction assay. African cassava mosaic virus (ACMV) and East African
cassava mosaic virus were detected on all the cultivars either as single infections or as
mixtures. The detection of EACMV on cassava at Fumesua and Ejura is the first to be reported
in Ashanti region in Ghana. This study recommended that, with the advent and spread of the
EACMV serotype of the mosaic virus in important cassava growing eco-zones and the
emergence of some severe strains of the African cassava mosaic in the pathosystem highly
resistant planting materials should be used for ratooning of mother plants as one of the
methods to increase the production of clean planting materials for farmers. It was also
indicated that, there is also the need to conduct an extensive survey in all the cassava growing
areas in Ghana to determine the incidence and spread of emerging species of the cassava
mosaic begomoviruses virus in order to develop better strategies to reduce the menace of the
cassava mosaic disease in Ghana and sub-region as a whole (Lamptey et al., 2011).

www.intechopen.com
Establishment of Functional Biotechnology Laboratories in Developing Countries 75

2.6 Genetic engineering

Genetic transformation, also referred to as Modern Biotechnology, is the application of the
techniques of molecular biology and/or recombinant DNA technology, or in vitro gene
transfer, to develop products or impart specific capabilities to organisms. Although the various
breeding programs in Ghana are still using conventional breeding tools, efforts are being made
to improve plants through an additional technique known as genetic engineering or
recombinant DNA (rDNA) science. This method does not rely on the pollination of flowers: it
allows individual genes with desire traits to be moved directly from one organism into another
living DNA of the same or different species. This technique is very vital for the improvement
of most of our local staple crops which are vegetatively propagated. Genetic engineering was
first accomplished in the laboratory in 1973 (Paarlberg, 2008). However, as of 2011, laboratories
in Ghana have not started applying rDNA in crop research activities. In 2008, a biosafety
legislative instrument was passed in the Ghanaian parliament to permit confined field trials
and contained laboratory experiment by research scientists. In 2009, a project to Strengthen
Capacity for the Safe Management of Biotechnology in Sub-Sahara Africa (SABIMA), was
initiated by the Forum for Agricultural Research in Africa (FARA) with sponsored by the
Syngenta Foundation for Sustainable Agriculture (SFSA). This project identified personalities
who can champion the advancement of Biotechnology in Ghana (Champions) and also focal
persons to run the project. Following advocacy and awareness creation workshops conducted
by champions in the SABIMA project, with focus on policy makers since 2010, the policy
makers were incited to pay critical attention to the biosafety bill that has been before
parliament. The members of parliament subsequently, took the biosafety bill through
consideration in parliament, in February 2011 and it was passed into law in June 2011. In
Ghana, there already exist a National biosafety committee (NBC) which has started receiving
applications for conducting confined field trials (CFT). The three proposals in the pipeline for
consideration by the NBC are:
1. Introduction of BT-Cowpea in Ghana
2. Protein quality improvement in sweetpotato
3. Nitrogen use efficiency and salt tolerance in rice
Other research efforts have been initiated in collaboration with advanced laboratories using
crops of local importance, and these include studies on the transgenic potential of Dioscorea
rotundata, using agrobacterium-mediated genetic transformation (Quain et al., 2011). This
study considered D. rotundata (yam) which is one of the important staples and sources of
carbohydrate in the diet in Sub-Saharan African sub-region. The crop has several post
harvest problems including poor storage of the tuber, availability of the edible planting
materials and high cost of labour for cropping. The crop therefore needs to be improved
using modern biotechnology methods. Studies were conducted to induce shoot regeneration
in D. rotundata leaf petiole as explants. Explants (0.5 cm long) were cultured on MS medium
supplemented with 0.2 mg L-1 2,4-D for 3 days, and transferred to MS medium containing
TDZ alone or in combination with 2iP. Shoot regeneration was observed within 21 days on
all media; however, the highest shoot regeneration, 7–9 shoots developing per explant were
obtained on media supplemented with TDZ and 2iP. The shoots grew vigorously when
transferred to MS medium supplemented with GA3. When petiole explants were subjected
to Agrobacterium-mediated transformation using strain C58 and EHA101 harbouring a
binary plasmid containing the β glucuronidase (gusA or uidA) intron gene under the
transcriptional control of CaMV35S promoter, very high efficiency of transformation (25–
65%) was obtained. This successful organogenesis and transformation protocol could be

www.intechopen.com
76 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

optimized and adapted in engineering of local yam quality and productivity for enhanced
protein content, and longer shelf-life.

2.7 Advancements achieved in Ghana

To facilitate research in the application of biotechnology tools, Ghanaian researchers have
partnered with regional, subregional and international programs. These include projects
with financial and technical support from International Atomic Energy Agency (IAEA),
Generation Challenge Program (GCP), United States Agency International Development
(USAID), CORAF/WECARD, Syngenta foundation for Sustainable Agriculture, United
States Department of Agriculture/Food and Agricultural Science (USDA/FAS), African
Agricultural Technology Foundation (AATF), United Nations University – Institute for
Natural Resources in Africa (UNU/INRA), World Bank, just to name a few. Collaboration
with CGIAR centers as well as universities (Tuskegee University Alabama, USA, and
Cornell University) has contributed immensely to recent research outputs, training human
capacity, as well as technical and infrastructural capacities.
Presently, Ghanaian agricultural research organizations have graduated to the status of
organizing national and regional training courses which hitherto were organized at
Consultative Group (CG) centers in the subregion. This activity re-enforces the fact that both
the human and infrastructural capacities have been developed to identify and solve regional
problems.
Development of functional and sustainable Biotechnology systems needs to take into
account `proper stewardship principle. Ghana is presently a participating country in the
project to Strengthen Capacity for Safe Application of Biotechnology (SABIMA). This is an
Excellence through stewardship based project ensuring the utilization of modern
biotechnology tools is practiced in a responsible manner. Under this project, through
stewardship training workshops, researchers are being trained to develop and properly
document standard operating procedures as well as critical control points along the life
cycle of product development. Biotechnology application policies are also being developed
by the various institutes for implementation at the management level in the organizations.
Over the years, Ghanaian Universities had been training plant and animal breeders and
researchers in related fields of applied and basic sciences, with limited or no knowledge in
molecular biology. There is therefore a generation of Breeders and Research Scientists with
little or no knowledge of molecular biology, tissue culture and modern biotechnology.
However with the establishment of the West Africa Centre for Crop Improvement (WACCI),
Alliance for Green Revolution in Africa (AGRA), Postgraduate Program at the University of
Ghana at Legon, and Kwame Nkrumah University of Science and Technology (KNUST)
respectively, a new generation of plant breeders with knowledge on the use of
biotechnology in crop improvement are being turned out. This new generation of plant
breeders is challenged to help solve African’s problems on African’s soil by developing crop
varieties tolerant/resistant to biotic and abiotic stresses, and thus alleviate poverty in order
to achieve the Millennium Challenge Goals. The Government of Ghana also assisted by
providing funds for the establishment of Biotechnology laboratories in 2006, the CSIR –
Crops Research institute and CSIR – Animal Research Institute. As a result, a new breed of
lecturers, researchers and students are using conventional and biotechnological tools for
crop and animal improvement. To us in the developing countries, some of the outmoded
technological tools in the advanced countries are novel ideas. The use of laborious and

www.intechopen.com
Establishment of Functional Biotechnology Laboratories in Developing Countries 77

conventional breeding methods with its attendant long duration have given way to
evaluation and selection using marker assisted breeding, mutation breeding, use of tissue
culture to select somaclonal variants and disease elimination, production of clean planting
materials, and mass production of clonal planting material by means of tissue culture.

3. Conclusion
The application of modern biotechnology in developing countries especially in Africa, has
great prospects. All the necessary efforts have to be employed in the form of financing,
policies, technologies, collaboration etc. These will help us to realize the inherent potentials
and immense contributions to the scientific advancement worldwide. All stakeholders are
needed to play their various roles to ensure the responsible application of biotechnology in
developing countries. The case of Ghana with respect to the CSIR-CRI alone stated above
gives the clear indication that, consistency, great leadership, team work, human and
infrastructural capacity building, good networking and collaboration are keys to
establishing a sustainable system. Presently, under the West African Agriculture
Productivity Program (WAAPP), with sponsorship from the World Bank, a multipurpose
biotechnology facility is being constructed to facilitate root and tuber research activities in
the sub-region. It is hopeful that the impetus will keep building up, and more innovative
strategies will be put in place to harness utilization of biotechnology tools in the sub-region.

4. Acknowledgement
The authors wish to acknowledge the following persons for the crucial roles they played in
training human resource and advocating for funds for the advancement of biotechnology in
Ghana, name; Dr. Elizabeth Acheampong, Dr. M. Egnin, Dr. C. Bonsi, Prof. P. Berjak, Dr,
Hans Adu-Dapaah, Prof. A Oteng-Yeboah, Dr. Y. Difie Osei, Prof. Walter Alhassan, Prof
Boampensem, Dr. J. Asafo-Adjei, just to mention a few. The CGIARs are also duly
acknowledged for their immense contribution in training human resource capacity, as well
as improving infrastructural capacity.

5. References
Acquah, W. E.; Quain M. D.; Twumasi, P. (2011). Genetic relationships between some released
and elite Ghanaian cassava cultivars based on distance matrices. African Journal of
Biotechnology Vol. 10(6), pp. 913-921, Available online at
http://www.academicjournals.org/AJB ISSN 1684–5315 © 2011 Academic Journals
Asibuo J. Y., He, G. ,Akromah, R.; Adu-Dapaah H.K. & Quain M.D. (2010). Genetic diversity of
groundnut botanical varieties using simple sequence repeats. .Aspects of Applied
Biology 96, Agriculture: Africa Engine for Growth- Plant science and biotechnology
the key.
Ashun, M. D. (1991) – Effect of various Hormonal (Growth regulators) combinations on in
vitro sprouting of various species of Dioscorea under light and dark conditions -
B.Sc. dissertation submitted to University of Ghana - Legon.
Ashun, M. D. (1996)- In vitro studies on micropropagation of various yam species (Dioscorea
species) M.Phil. Thesis submitted to University of Ghana - Legon.
Benson E.E., & Bremner, D. (2004). Oxidative stress in the frozen plant: a free radicle point of
view. In: B.J. Fuller N. Lane E.E. Benson (eds). Life in the frozen state, CRC Press Boca
Raton. pp 205 – 241.

www.intechopen.com
78 Biotechnology - Molecular Studies and Novel Applications for Improved Quality of Human Life

Lamptey, J.N.L.; Quain, M.D.; Owusu Kyere, E.; Ribeiro,P.F.; Prempeh,R.; Afriyie-Debrah,
C. & Abrokwa,L. (2011). Effect of Cassava Mosaic Disease on Successive Ratooning
of Some Cassava Cultivars. The West Africa Root and Tuber Crops Conference,
Abstract, pp 10.
Miller, S.A.; & Martin. R.R.; (1988) Molecular Diagnosis of Plant Disease. Ann. Rev. Phytopathol.
26 pp 409-32Molecular Biology Source
http://en.wikipedia.org/w/index.php?oldid=417169395
Murashige, T. & Skoog, F. (1962) A revised medium for rapid growth and bioassays with
tobacco tissue cultures. Physiol. Plant. 15:473-497
Murch, S.J. & Saxena, P.K. (Eds.). (2005). Journey of a Single Cell to a Plant. Oxford & IBH
Publishing Co. Pvt. Ltd., New Delhi, India
Otoo, J. A. & Quain, M. D. (2001) comparative studies of in vitro cleaned planting material
and field selected apparently clean sweet potato planting material in proceedings
of 8th International Society for Tropical Root crops - Africa Branch (ISTRC-AB)
symposium at Cotonou 2001.
Paarlberg (2008). Starved being for Science How Biotechnology is being kept out of Africa. ISNB-
13:978-0-674-02973-6
Panis B., & Lambardi, M. (2005), Status of Cryopreservation Technologies. In Plants (Crops and
Forest Trees). In The Role of Biotechnology Villa Gualino, Turin, Italy–5-7 March, pp. 43–54.
Quain, M.D.; Egnin, M.; Bey, B.; Thompson, R. & Bonsi, C. (2011). Transgenic potential of
Dioscorea rotundata, using agrobacterium-mediated genetic transformation. Aspects
of Applied Biology 110, ISSN 0265-1491 pp71-79
Quain M. D., Adofo-Boateng, P.; Mensah Dzomeku, B.; & Agyeman, A. (2010a). Multiple
Shoot Generation Media for Musa sapientum L. (False Horn, Intermediate French
Plantain and Hybrid Tetraploid French Plantain) Cultivars in Ghana. The African
Journal of Plant Science and Biotechnology. 4 (Special Issue 1), 102 – 106, © 2010 Global
Science Books.
Quain, M. D.; Thompson, R.; Omenyo, E.L.; Asibuo, J.Y.; Appiah-Kubi, D.& Adofo-Boateng,
P. (2010b). Assessing transferability of Sweetpotato EST SSR primers to cocoyam
and micropropagation of nine elite cocoyam varieties in Ghana. Aspects of Applied
Biology 96, Agriculture: Africa Engine for Growth - Plant science and biotechnology
the key. pp. 269-276.
Quain, M. D.; Dzomeku, B.M.; Thompson,R.; Asibuo, J.Y.; Boateng, P.A. & Appiah-Kubi,
D. (2010c). Genetic Diversity of Musa cultivars in Ghana and introduced hybrids.
Aspects of Applied Biology 96, Agriculture: Africa Engine for Growth- Plant science
and biotechnology the key. pp. 277-28
Quain, M. D. (2001.) Propagation of Root and Tuber Crops By Tissue Culture In Ghana. – West
Africa Seed and Planting Material Newsletter of the West Africa Seed Network (WASNET).
Issue No. 8
Quain, M. D. & Acheampong, E. (2001). Effect of Explant Age on In vitro Development of
three Dioscorea species. Proceedings of 7th International Society for Tropical Root crops
- Africa Branch (ISTRC-AB) symposium at Cotonou 2001.
Ribaut, J.M. & Hoisington, D. A. (1998). Marker assisted selection: new tools and strategies.
Trends Plant Sci., 3, 236–239
Secretariat of the Convention on Biological Diversity (2000). Cartagena Protocol on Biosafety
to the Convention on Biological Diversity: text and annexes. Montreal:Secretariat of
the Convention on Biological Diversity. ISBN: 92-807-1924-6
Walters, C.; Hiu, L.M. & Wheeler, L.M. (2005). Dying while dry: kinetics and mechanisms of
deterioration in desiccated organisms. Integrative Comparative Biology 45, pp. 751-758.

www.intechopen.com
 Biotechnology - Molecular Studies and Novel Applications for
Improved Quality of Human Life
Edited by Prof. Reda Sammour

ISBN 978-953-51-0151-2
Hard cover, 250 pages
Publisher InTech
Published online 14, March, 2012
Published in print edition March, 2012

This book deals with the importance of application of molecular biology as an approach of biotechnology for
improvement of the quality of human life. One of the interesting topics in this field, is the identification of the
organisms that produce bioactive secondary metabolites. It also discusses how to structure a plan for use and
preservation of those species that represent a potential source for new drug development, especially those
obtained from bacteria. The book also introduces some novel applications of biotechnology, such as
therapeutic applications of electroporation, improving quality and microbial safety of fresh-cut vegetables,
producing synthetic PEG hydro gels to be used as an extra cellular matrix mimics for tissue engineering
applications, and other interesting applications.

How to reference
In order to correctly reference this scholarly work, feel free to copy and paste the following:

Marian D. Quain, James Y. Asibuo, Ruth N. Prempeh and Elizabeth Y. Parkes (2012). Establishment of
Functional Biotechnology Laboratories in Developing Countries, Biotechnology - Molecular Studies and Novel
Applications for Improved Quality of Human Life, Prof. Reda Sammour (Ed.), ISBN: 978-953-51-0151-2,
InTech, Available from: http://www.intechopen.com/books/biotechnology-molecular-studies-and-novel-
applications-for-improved-quality-of-human-life/establishment-of-functional-biotechnology-laboratories-in-
developing-countries

InTech Europe InTech China

University Campus STeP Ri Unit 405, Office Block, Hotel Equatorial Shanghai
Slavka Krautzeka 83/A No.65, Yan An Road (West), Shanghai, 200040, China
51000 Rijeka, Croatia
Phone: +385 (51) 770 447 Phone: +86-21-62489820
Fax: +385 (51) 686 166 Fax: +86-21-62489821
www.intechopen.com
© 2012 The Author(s). Licensee IntechOpen. This is an open access article
distributed under the terms of the Creative Commons Attribution 3.0
License, which permits unrestricted use, distribution, and reproduction in
any medium, provided the original work is properly cited.